Kidney Week

Abstract: FR-OR44

Single-Cell Profiling of Peripheral Blood Mononuclear Cell Identifies Immune Populations Associated with High Risk of Early Acute Rejection

Session Information

• In-Depth Look at Transplantation: Basic and Translational
  October 23, 2020 | Location: Simulive
  Abstract Time: 05:00 PM - 07:00 PM

Category: Transplantation

• 1901 Transplantation: Basic

Authors

• Zhang, Weijia, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Weijia Zhang

• Farouk, Samira S., Icahn School of Medicine at Mount Sinai, New York, New York, United States

Samira S. Farouk,

• Sun, Zeguo, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Zeguo Sun,

• Yi, Zhengzi, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Zhengzi Yi,

• Fu, Jia, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Jia Fu,

• Wei, Chengguo, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Chengguo Wei,

• Fribourg, Miguel, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Miguel Fribourg,

• He, John Cijiang, Icahn School of Medicine at Mount Sinai, New York, New York, United States

John Cijiang He,

• Cravedi, Paolo, Icahn School of Medicine at Mount Sinai, New York, New York, United States

Paolo Cravedi,

• Murphy, Barbara T., Icahn School of Medicine at Mount Sinai, New York, New York, United States

Barbara T. Murphy,

Background

Our previous transcriptomic analysis of pre-transplant peripheral blood of 235 kidney transplant recipients in a multi-center GoCAR cohort revealed a 23-gene set predictive of early acute rejection (EAR) post kidney transplant, but the associated immune profile is unknown.

Methods

We first developed a reliable targeted RNA expression (TREx) sequencing assay based on the EAR prediction gene set and applied the assay to assess the EAR risk of 21 dialysis patients who were recruited at Mount Sinai Hospital. We next performed extensive singe-cell immune profiling of PBMCs of these patients using 33 surface markers with CyTOF technology. Lastly, using 10X genomics technology, we performed single cell RNA sequencing (scRNAseq) of the PBMCs to identify immune cell (sub)populations and their transcriptomic signatures from two patients with high and low risk of EAR, respectively.

Results

Using the established TREx assay to stratify patients, we identified had 6 high, 9 intermediate and 6 low risk patients. CyTOF immune profiling of PBMCs of these patients indicated that NK cell population was negatively associated with the risk score (p=0.008) while CD4 T cells were positively associated (p=0.056). scRNAseq analysis of the transcriptomic profiles of a total number of 11,966 cells confirmed these findings. In addition, scRNA sequencing profiling identified NK and T cell subsets and their transcriptomic signatures associated with the EAR risk, especially decreased NK CD56^Dim and increased CD4 CTL populations in EAR high risk patients, which was further validated using PBMC composition deconvoluted from bulk RNAseq profiles of GoCAR cohort (n=235).

Conclusion

The Baseline Acute Rejection assay is associated with clear cellular and molecular Phenotype which may help us further understand the underlying mechanisms that lead to the development of acute rejection.